Off-season flowering in red pitaya (Hylocereus sp.), a long-day plant, can be achieved using night-breaking (NB) treatment. Among the stages of bud development, stage 0 referred to induced but not yet differentiate any bracteole and stage 3 was the stage right before emerging floral buds and the bracteole differentiation was completed. Unlike floral bud emergence, bracteole differentiation was independent of the daylength and strongly influenced by the environmental temperature. The buds of higher stages were more effective in response to NB treatment and more sensitive to chilling injury (CI). Consequently, off-season flowers in autumn and winter trials were derived mainly from stage 2 and 3 buds and from stage 0 and 1 buds, respectively. In southern Taiwan, low night temperature between 10 Jan. and 7 Feb. 2011 may be the major factor, which delay bud development in off-season production. Therefore, we conducted a heating experiment in winter off-season production to proof our hypothesis and concluded that NB treatment should be applied along with night temperature elevation or after mid-February when the minimum night temperature is increasing.